H3K79me1 Polyclonal Antibody

$Western blot was performed on histone extracts from HeLa cells (15 \u03bcg) with the H3K79me1 antibody (bs-53049R), diluted 1:1,000 and 1:2000 in TBS-Tween containing 5% skimmed milk. The molecular weight marker (Bio-Rad, broad range biotinylated SDS-PAGE standard) is shown on the left, the location of the protein of interest is indicated on the right.$

Western blot was performed on histone extracts from HeLa cells (15 \u03bcg) with the H3K79me1 antibody (bs-53049R), diluted 1:1,000 and 1:2000 in TBS-Tween containing 5% skimmed milk. The molecular weight marker (Bio-Rad, broad range biotinylated SDS-PAGE standard) is shown on the left, the location of the protein of interest is indicated on the right.

A Dot Blot analysis was performed to test the cross reactivity of the H3K79me1 antibody (bs-53049R) with peptides containing other modifications of histone H3. These include di- and trimethylation of the same lysine and mono-, di- and trimethylation of lysine 9, 27 and 36. One hundred to 0.2 pmol of the peptides were spotted on a membrane. The antibody was used at a dilution of 1:100,000. The figure shows a high specificity of the antibody for the modification of interest.

To determine the titer of the antibody, an ELISA was performed using a serial dilution of the H3K79me1 antibody (bs-53049R) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:30,000.

$ChIP assays were performed using HeLa cells, H3K79me1 antibody (bs-53049R) and optimized PCR primer pairs for qPCR. Chromatin from 1.6 million cells were sheared. ChIP was performed with IgG (5 \u03bcg\/IP) as a negative IP control. The IP\u2019d DNA was analysed by qPCR using primers for different positive and negative loci. The results are expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). The figure shows recovery of the GAPDH promoter and myoglobin exon 2 with a titration of the H3K79me1 antibody consisting of 5, 10 and 15 \u03bcl per ChIP experiment.$

ChIP assays were performed using HeLa cells, H3K79me1 antibody (bs-53049R) and optimized PCR primer pairs for qPCR. Chromatin from 1.6 million cells were sheared. ChIP was performed with IgG (5 \u03bcg\/IP) as a negative IP control. The IP\u2019d DNA was analysed by qPCR using primers for different positive and negative loci. The results are expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). The figure shows recovery of the GAPDH promoter and myoglobin exon 2 with a titration of the H3K79me1 antibody consisting of 5, 10 and 15 \u03bcl per ChIP experiment.

$ChIP assays were performed using HeLa cells, H3K79me1 antibody (bs-53049R) and optimized PCR primer pairs for qPCR. Chromatin from 1.6 million cells were sheared. ChIP was performed with IgG (5 \u03bcg\/IP) as a negative IP control. The IP\u2019d DNA was analysed by qPCR using primers for different positive and negative loci. The results are expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). The figure shows recovery of RPL30, ALDOA, SERPINA1 and SAA4 using 10 \u03bcl of antibody per ChIP experiment.$

ChIP assays were performed using HeLa cells, H3K79me1 antibody (bs-53049R) and optimized PCR primer pairs for qPCR. Chromatin from 1.6 million cells were sheared. ChIP was performed with IgG (5 \u03bcg\/IP) as a negative IP control. The IP\u2019d DNA was analysed by qPCR using primers for different positive and negative loci. The results are expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). The figure shows recovery of RPL30, ALDOA, SERPINA1 and SAA4 using 10 \u03bcl of antibody per ChIP experiment.

Applications

WB
ELISA
dot-blot

Reactivity

Human

Overview
Catalog #	bs-53049R
Product Name	H3K79me1 Polyclonal Antibody
Applications	WB, ELISA, dot-blot
Reactivity	Human
Specifications
Conjugation	Unconjugated
Host	Rabbit
Source	Polyclonal antibody raised in rabbit against histone H3 the monomethylated lysine 79 (H3K79me1), using a KLH-conjugated synthetic peptide
Clonality	Polyclonal
Isotype	Not Determined
Concentration	Not Determined
Purification	Whole antiserum from rabbit containing 0.05% azide
Storage Buffer	Serum with 0.05% sodium azide.
Storage Condition	Store at -20°C for 12 months.
Target
Gene ID	8350
Swiss Prot	P68431
Background	Core component of nucleosome. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling.
Application Dilution
WB	1:300-5000
ELISA	1:500-1000
dot-blot